Matrix mixing chamber

Abstract

The invention discloses mixing equipment for reconstructing a powdery cell culture medium. The apparatus includes at least one fluid chamber and an inflow port at a lower portion of the fluid chamber. The apparatus also includes a geometric fluid flow aid located in the fluid chamber and an outflow port located at the top of the fluid chamber. Powdery cell culture medium is provided in the fluid to mix with the fluid provided by the inflow port. The outflow port is configured to allow the reconstructed substrate to exit the fluid chamber.

Description

[0001] This application is a divisional application of the patent application with application number 201680078727.9 (PCT / US2016 / 046603) filed on August 11, 2016 and invention name “Matrix Mixing Chamber”.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This patent application claims priority to U.S. patent application serial number 15 / 087,826 filed on March 31, 2016, which claims priority to U.S. provisional application serial number 62 / 257,685 filed on November 19, 2015, the entire contents of each of the above applications are incorporated into this application by reference. Technical Field

[0004] Embodiments of the present technology generally relate to mixing devices for mixing at least one input fluid stream with a material to be mixed with one or more input fluid streams. More specifically, embodiments of the present technology relate to mixing devices particularly suitable for reconstituting powdered cell culture media in predetermined unit volume amounts. Background Art

[0005] Animal cells and tissues that can survive in vitro culture have been known since the early 20th century. Although animal cell culture today is a sophisticated technology, the basic culture techniques have not changed since the beginning of this century. Native or transformed cells or tissues are grown in a liquid nutrient mixture often referred to as a "matrix." For example, the matrix can be a complex mixture of amino acids, vitamins, salts, and other ingredients. It is usually supplemented with 1-10% purified fetal bovine or newborn calf serum. Cell culture media and serum can be obtained commercially from many sources.

[0006] While basic cell culture techniques have not changed significantly over the years, cell culture volumes and accessibility to laboratory technology continue to increase dramatically, changing the demands on cell culture technology. Not only are more research labs, pharmaceutical, and biotech companies adopting tissue culture technology, they are often doing so on a relatively large scale. Medical product companies may consume tens or hundreds of liters of liquid matrix per day and employ large numbers of lab technicians and scientists to produce antibodies, growth factors, or purified proteins from tissue culture for commercial use. As a result, between matrix supply costs and employee time, there are considerable expenses associated with today's tissue culture processes.

[0007] Cell culture media are usually commercially available as dry powders that are reconstituted by adding an appropriate volume of water or as prepackaged liquids. There are also a variety of additives that are usually added to the medium before use. These additives include sodium bicarbonate, glutamine, additional buffers or antibiotics.

[0008] The technology described herein generally relates to improved devices and methods for mixing substrates in various volumes and environments. Summary of the invention

[0009] In general, embodiments described herein relate to apparatus and methods for powdered matrices that are easy to prepare, require less storage space than liquid matrices, and / or require minimal effort to prepare. The technology according to some embodiments relates to a mixing device for mixing a material (e.g., a powdered matrix) with at least one input fluid stream. More specifically, some embodiments of the technology relate to mixing devices particularly suitable for reconstituting powdered cell culture media in predetermined unit volume amounts and various methods related thereto.

[0010] The present technology represents a significant improvement in this area of ​​technology.

[0011] Some embodiments are at least partially based on some defects and / or inconveniences of existing substrate technology as recognized by the inventor of the prior art, or based on the understanding to the inventor's potential improvement.For example, pre-packaged liquid can be aseptic and be divided into convenient size, and can be ready to use.Yet this substrate is usually light sensitive and has the shelf life of regulation.Therefore, substrate must be ordered regularly.It also should be stored refrigerated, and its pre-packaged form, needs a large amount of manpower time to unpack and transport.In addition, the freight cost of pre-packaged liquid becomes more and more expensive.

[0012] In addition, powdered substrates are provided in batches or with pre-weighed packaging. It tends to have a longer shelf life, cheaper and requires less storage space and processing time than liquid forms. However, powdered substrates must be dissolved and divided equally under aseptic conditions. Especially for large volume substrate preparation, although cost increases, the increased processing and preparation time usually make pre-packaged liquid substrates become preferred.

[0013] The reconstruction of powdered matrix is ​​usually a several step process. In order to prepare a liquid matrix from a solid powder, a known amount of powder for a specific volume of matrix is ​​measured and added to a certain volume of distilled water, which is usually slightly less than the final desired volume. The powder and water are stirred until the solid is completely dissolved. A specific amount of sodium bicarbonate is added and dissolved. Thereafter, an acid or alkali can be used to adjust the pH, and additional water is added to increase the matrix to its final volume. The whole mixture is then passed through a sterilizing filter. Subsequently, the matrix can be collected in a single larger sterile container, or distributed in several smaller sterile containers.

[0014] Powdered tissue culture medium has a very fine particle size and is hygroscopic. It has a tendency to "ball" or "clump" when mixed with water. Therefore, when reconstituted in water or other aqueous liquids, sufficient stirring is required to break up any clumps that may form upon initial contact with water. For smaller batches, a sterile magnetic stir bar can be added to the mixing vessel, which is then placed on a magnetic stir plate. Additional manipulation is usually required to add the stir bar to the mixing vessel. In a typical laboratory setting, a magnetic stir plate is not a practical solution for large volume matrix preparation.

[0015] Additionally, due to its hygroscopic nature, the matrix absorbs moisture when stored, especially in a humid environment. Wet matrix has a shortened shelf life, becomes lumpy, and requires active agitation to reconstitute. Therefore, the powdered matrix shelf life can be improved if it is provided in pre-weighed, sealed, and dried aliquots.

[0016] The reconstitution process requires several steps and several separate parts of equipment. It usually requires at least one container large enough to accommodate the entire final volume of the reconstitution matrix, plus one or more containers to receive the sterile matrix after filtration. The sterilized matrix is ​​usually delivered to a top-opening container. Therefore, most of the matrix preparation is completed in a laminar flow hood. Since there is usually not enough space to accommodate containers and sterile matrix, it is difficult to handle large-volume matrix in the hood. Some embodiments herein are based on such recognition that a device that allows the preparation of large-volume products with minimal physical contact and is convenient for matrix preparation without the above-mentioned inconvenience will meet the long-term unmet needs of the scientific community.

[0017] Furthermore, the equipment used to reconstitute the powdered matrix must be thoroughly cleaned between uses to remove residues and eliminate contaminants in the equipment. This requires a great deal of time, and even with careful work, the risk of contaminants remaining remains. Therefore, some of the following embodiments relate to a low-cost equivalent of a matrix mixing chamber that is constructed for single use.

[0018] One embodiment of the technology relates to a mixing device for reconstructing a powdered cell culture medium. The device includes at least one fluid chamber, a first inlet port located at the top of the fluid chamber, and a second inlet port located at the bottom of the fluid chamber. The device also includes a geometric fluid flow aid positioned in the fluid chamber and an outflow port located at the top of the fluid chamber. The powdered cell culture medium is provided in the fluid to mix with the fluid provided by at least one of the first inlet port or the second inlet port. The outflow port is configured to allow the reconstructed matrix to leave the fluid chamber.

[0019] A second embodiment of the present technology relates to a method for reconstructing a powdered cell culture medium. The method includes providing a mixing device having at least one fluid chamber, a first inlet port located at the top of the fluid chamber, a second inlet port located at the bottom of the fluid chamber, a geometric fluid flow aid positioned in the fluid chamber, and an outflow port located at the top of the fluid chamber. The method also includes providing the powdered cell culture medium to the fluid chamber before introducing the fluid into the chamber, and then introducing the fluid into the chamber through at least one of the first inlet port and the second inlet port. The first inlet port and the second inlet port are oriented tangentially relative to the inner wall of the fluid chamber so that the fluid moves along the wall of the fluid chamber in a circular motion. The method further includes generating a vortex in the fluid chamber by introducing the fluid at a sufficient flow rate, and enhancing the vortex by further directing water around the wall of the fluid chamber around the geometric flow aid using the geometric fluid flow aid. The method includes continuing to introduce the fluid into the chamber and collecting the reconstructed fluid that leaves the chamber through the outflow port. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] The above-mentioned features and other features, aspects and advantages of the present technology will now be described in conjunction with various embodiments of the present invention and with reference to the accompanying drawings. However, the illustrated embodiments are only exemplary and are not intended to limit the present invention.

[0021] Figure 1 is a schematic diagram of an integrated mixing chamber, sterilizing filter, and receiving vessel system according to one embodiment.

[0022] Figure 2 yes Figure 1 A cross-sectional view of an embodiment of a mixing chamber.

[0023] Figure 3 is an exploded view of a mixing chamber according to a second embodiment.

[0024] 4 is a cross-sectional view of a mixing chamber with a representation of a fluid vortex according to the prior art.

[0025] 5a is a top perspective view of a top cone of a mixing chamber according to one embodiment.

[0026] 5b is a bottom perspective view of the top cone shown in FIG. 5a.

[0027] Figure 6 is a top perspective view of a bottom cone of a mixing chamber according to one embodiment.

[0028] Figure 7 is a diagram showing an assembly of a bottom cover and Figure 6 A top perspective view of the lower cover of the mixing chamber of the bottom cone shown in FIG.

[0029] 8a, 8b and 8c are perspective views of the bottom of the middle cover showing the assembly of the middle cover and the mixing chamber of the top cone shown in Figs. 5a and 5b according to one embodiment.

[0030] 9a, 9b, 9c, and 9d are perspective views of the bottom of a top cover of a mixing chamber showing the assembly of the top cover according to one embodiment.

[0031] Fig.10 is a perspective view of a cap attached to an outlet tube according to one embodiment.

[0032] Fig.11 is a perspective view of a bottom cover attached to an inlet tube according to one embodiment.

[0033] Fig.12 is a perspective view of an intermediate cover attached to an inlet tube and a pressure sensor according to one embodiment.

[0034] Fig.13 is a schematic diagram of a single chamber mixing device according to one embodiment. DETAILED DESCRIPTION

[0035] In the following detailed description, reference is made to the accompanying drawings that form a part of the present disclosure. In the drawings, similar symbols generally designate similar parts unless the context dictates otherwise. The illustrative embodiments described in the detailed description, drawings, and claims are not meant to be limiting. The detailed description is intended as a description of exemplary embodiments and is not intended to represent the only embodiment that can be practiced. The term "exemplary" as used herein means "used as an example, instance, or illustration" and is not necessarily interpreted as being preferred or advantageous over other embodiments. Other embodiments may be used, and other changes may be made without departing from the spirit or scope of the subject matter presented herein. It will be readily understood that aspects of the present disclosure, as generally described herein and shown in the drawings, may be arranged, replaced, combined, and designed in a variety of different configurations, all of which are expressly contemplated and form a part of the present disclosure.

[0036] Embodiment described herein generally relates to storage, transportation, preparation and / or use relevant device / equipment, system and method with matrix, for example, the matrix for cell culture.One or more in the embodiment provided can overcome one or more in the shortcoming, restriction or defect existing in the prior art, particularly about the restriction of matrix box and this box, comprise those boxes with dry powder matrix.For example, in some embodiments, device / equipment can be single use, disposable or preloaded with the material of expectation.These devices can comprise one or more parts in material mixing and matrix use in allowing to improve asepticity, storage duration, transportation, device.The system and method of making and using these devices have also been described herein.

[0037] Figure 1 is an overall system view of one embodiment of a mixing device 10, a filter 58, and a receiving bag 50, according to certain embodiments. Figure 2 1 is a cross-sectional view showing an internal view of components of the mixing device 10. The mixing device 10 includes at least one chamber, and in some embodiments, two chambers. The generally cylindrical first chamber 12 constitutes the lower chamber in the preferred embodiment depicted herein, and the second chamber 14 constitutes the upper chamber of the preferred embodiment. For descriptive purposes, in the dual chamber embodiment, "chemical A" will refer herein to the material contained in the first chamber 12, and "chemical B" will refer to the material contained in the second chamber 14.

[0038] The input fluid stream enters the mixing device 10 through at least one inlet port. It is contemplated that although the present application will describe embodiments having multiple inlet ports, for example Figure 1 In the embodiment shown in FIG. 1 , two inlet ports 16a, 16b are shown, but the features of the mixing device 10 described herein can still be applied to a mixing device 10 having one inlet port. The axis of the one or more inlet ports relative to the first chamber 12 can be such that the fluid flow enters substantially at a tangent angle relative to the inner wall of the first chamber 12, so that the fluid entering the first chamber through the inlet ports 16a, 16b along the side of the chamber to form a circular mixing motion that helps the mixing of chemical A with the fluid flow in the first chamber. The use of more than one inlet port helps to direct the flow toward the undissolved volume of chemical A, which is usually in the form of a solid powder. The upper inlet port 16b also helps prevent the solid matrix from adhering to the top of the first chamber 12. The inlet ports 16a, 16b can be used simultaneously, or can be used alternately to achieve the necessary level of fluid movement to mix chemical A with the fluid flow.

[0039] As Chemical A dissolves in the liquid and additional liquid enters the first chamber 12, the liquid level advances upward through the middle cover 31 and into the second chamber 14. The fluid containing Chemical A that passes through the middle cover 31 and into the upper chamber comes into contact with Chemical B.

[0040] In preferred embodiments, chemical B has increased solubility properties over chemical A, such that no significant agitation is required to promote dissolution of chemical B in a liquid already containing chemical A.

[0041] Thereafter, the liquid containing dissolved chemicals A and B is preferably filtered through filter 48 ( Figure 3 , 9B ) then leaves the second chamber 14 through the upper cover 41 through the outflow port 49. In a preferred embodiment, the liquid (being Figure 1The tube shielding in Figure 2 ) enters sterilizing filter 58. Sterile filters of the type contemplated by the present technology can be purchased from many suppliers. An example of a commercial supplier is Pall Corporation, Courtland, ME. For sterile matrix products, the sterilizing filter device typically includes a 0.2μ filter. The filter may include nylon or cellulose acetate. Thereafter, the sterile liquid containing chemical A and chemical B leaves filter 58 and enters receiving container 50. In some embodiments, a hydrophobic vent filter is used between the outflow port and sterilizing filter 58 to exhaust air entrained in the dissolved medium so that it does not fill the sterilizing filter.

[0042] Additionally, it is contemplated that other types of filter sizes may be selected for other functions. For example, preparation of electrophoresis buffer requires a clean, but not necessarily sterile solution, so a 0.45μ filter may be sufficient. Similarly, preparation of a more viscous solution may require a wider pore size. In short, the filter may be of any desired size, volume, pore size, etc., to suit the particular use of the cartridge. For other applications of the technology disclosed herein, no additional filtering equipment is required. The liquid then passes directly to the receiving container via any suitable fluid communication means (e.g., flexible tubing). If a sterile filter is used, the tubing and all additional chemicals entering the multiple inlet ports 52 and the receiving container 50 should be sterile (see Figure 1 ).

[0043] It is further contemplated that the final product may require the addition of one or more other liquid additives, or that the container 50 may discharge into a series of different receptacles. Thus, one or more inlet / outlet ports, generally designated as a plurality of inlet / outlet ports 52, are typically provided. A stop flow regulator 54 is preferably associated with each of the inlet ports to provide control over the sequential discharge or inflow of the desired additive solution.

[0044] Figure 3 An exploded view of an embodiment of the mixing device 10 is described in detail. The mixing device base or lower cover 21 is combined with the first chamber housing 22 in association with a seal 23. A support structure 24 is preferably integrally molded with the lower cover 21 or milled into the lower cover 21 to form a shoulder or lip to support the first chamber housing 22. The support structure 24 and the first chamber housing 22 are preferably substantially cylindrical to optimize the rotational speed of the fluid that has been driven under pressure through the inflow port. The seal 23 is preferably an elastomeric O-ring, but can be a gasket or other sealing device known to those skilled in the art. The bottom cover also supports a geometric flow aid, such as a bottom cone 25. The components of the lower cover 21 will be referred to below. Figure 7 Describe in more detail.

[0045] The first chamber housing 22 is provided with one or more inlet ports 16 (at Figure 1 2) receives the inflow fluid, and these inflow ports 16 are generally tangentially oriented relative to the inner wall of the shell. One or more inflow ports 16 can be integrally molded with the shell 22, or can be fixed to the shell in any of various ways known in the art, such as by adhesives, solvents or thermal bonding techniques. According to the embodiment shown, two inflow ports 16 are integrally molded with the lower cover 21 and the middle cover 31 having a channel, wherein the channel allows fluid flow between the inflow port 16 and the interior of the shell 22. Hose barbs or other conventional connectors are preferably fixed to the inflow ports 16. Preferably, the inflow port 16a is located in the lower half of the first chamber 12, and more preferably along the lower quarter of the first chamber 12, such as on the lower cover 21. Preferably, the inflow port 16b is located in the upper half of the first chamber 12, and more preferably along the upper quarter of the first chamber 12, such as on the middle cover 31. In embodiments with a single inflow port, the inflow port can be located in the lower half of the first chamber 12, and more preferably along the lower quarter of the first chamber 12, such as on the lower cover 21. A protective cover can be provided to cover one or more inflow ports to prevent powder from spilling prior to use. In some embodiments, a quick-connect device is employed at the inflow port such that the quick-connect device prevents the powdered cell culture medium from detaching from the cartridge during storage and shipping.

[0046] Fluid entering the second port at sufficient velocity contributes to the vortex created by the fluid entering from the first port. For reconstitution of large quantities of dry powders or viscous solutions, two inlet ports may better promote complete mixing. Thus, water or other solvents may be added from more than one inlet port simply to support vortex generation. Alternatively, the liquids entering the device through multiple inlet ports may have different chemical compositions.

[0047] like Figure 1 As shown, the inlet ports can be positioned on the same vertical plane or along different vertical planes depending on the specific requirements of a given application, as long as the inlet flow from port 16a does not interfere with the inlet flow from port 16b. The fluids entering the mixing chamber tangentially from the two ports should flow in concert to maintain vortex activity.

[0048] It is conceivable that inflow ports 16a and 16b have equal port diameters. However, for different inflow flow rates, the diameter can be modified separately. The internal diameter and inflow pressure of each port can change to promote the mixing of required reagents. The smaller diameter port located above the larger diameter port will provide additional inflow velocity than the larger diameter port. In this way, effective eddy currents can be maintained to maximize the reconstruction of a given powder mixture. These design features will be added or included according to the solubility of the powder in a specific application, the volume of the powder relative to the chamber size and by the chemical properties required for the reconstruction of a given liquid formulation.

[0049] In use, liquid enters the mixing chamber through inflow ports 16a, 16b. Tap pressures or other inflow pressures exceeding about 1psi are generally strong enough to allow proper device function. For many embodiments, a typical interface pressure in the range of about 25psi is sufficient. As will be appreciated by those skilled in the art, the minimum effective pressure is a function of the size of the first mixing chamber, the volume of chemical A contained therein, and the diameter of the inflow lumen. Some routine experiments may need to optimize these parameters for a particular application. For example, these parameters can be designed to be specific to an available water source. When the source fluid pressure cannot be increased, the present mixing device can be designed by reducing the diameter of the inlet.

[0050] As previously mentioned, liquid enters the first chamber under pressure in a manner that is substantially tangential to the chamber inner wall. The speed of the liquid entering the device is determined by the pressure of the input fluid stream, and can be additionally manipulated by changing one or more diameters of one or more inflow ports or the size of the first chamber. The reduced inflow port diameter will increase the speed of the liquid entering the chamber, and the increased inflow port diameter will reduce the liquid speed. In a preferred embodiment, when the ratio of the cross-sectional area (in inches) of the inflow port through which the fluid enters the chamber to the flow rate (liters / minute) is 0.0015-0.0040, sufficient fluid speeds are achieved. In some preferred embodiments, the ratio is 0.0018 to 0.0038. In some preferred embodiments, the ratio is 0.0028+ / -0.0002 (in other words, 0.0028 square inches for the inlet of 1 liter of fluid flow per minute).

[0051] It is contemplated that for proper function of the mixing chamber for other applications, slight modifications to the apparatus will be required. For example, if the liquid is water and the product is tissue culture medium, normal tap pressure matched to the appropriate inlet port size will produce sufficient liquid pressure to produce the desired rotating fluid velocity. The mixing chamber inlet port diameter has a direct effect on the inlet velocity. As described above, the inlet diameter can be increased or decreased to adjust the velocity in order to provide adequate mixing of the substrate.

[0052] In the embodiment with two inflow ports (for example 16a and 16b), preferably at first fluid is provided to the chamber by upper inflow port 16b, to prevent undissolved powdered matrix agglomerates from floating. Once fluid has partially filled the chamber, lower port 16a opens and begins to promote vortex-shaped fluid flow motion.

[0053] Preferably, the pressure of the liquid stream combined with the compatible inflow port diameter will provide enough liquid velocity so that the liquid entering the device continues to travel along the surface of the inner chamber housing and along a circular path toward the center of the chamber. If the rotating fluid velocity of the liquid is sufficient, the motion then establishes a turbulent vortex, which is used to mix the inflow liquid with the contents of the first chamber. This motion is shown in Figure 4, representing the desired fluid motion in the prior art mixing device. The horizontal dotted line 80 represents the vortex fluid that forms a roughly conical air area 82 at its center. The vortex vortex mixes the contents of the chamber. The additional fluid entering the chamber pushes the vortex along the side of the chamber and toward the top. Although this vortex motion occurs under the right conditions, it is not always possible to achieve this now as the powdered solid chemicals change over time. In many cases, due to the current matrix, the vortex does not occur, and the incoming fluid is just injected into the chamber. In these cases, if there is no proper stirring, the powdered matrix will contact the fluid, but it will not be completely wetted to dissolve in the fluid. Agglomerates of the powdered matrix then float, adhere to surfaces of the chamber, and / or block inflow or outflow ports in the chamber.

[0054] Thus, embodiments of the mixing device 10 according to some embodiments assist in achieving and enhancing the vortex motion of the fluid by including one or more geometric flow aids (such as one or more cones along the contours of the bottom cone 25 and the upper cone 35) within the volume of the first chamber 12. It should be understood that, although multiple cones are depicted, some embodiments contemplate a single cone flow aid, while in other embodiments, additional aids may be included. Thus, as the fluid flows into the chamber via the inlet port 16, the fluid is directed into the chamber so that a vortex eddy is immediately generated to establish turbulent fluid motion for mixing the inflowing fluid with the contents of the first chamber 12. In addition, the geometric flow aid further enhances the mixing of the solid contents with the inflowing fluid by dispersing the solids and preventing the solids from adhering or agglomerating at the bottom or top of the first chamber 12. The geometric flow aid preferably includes a geometric shape having a decreasing diameter in a direction toward the center of the first chamber 12, thereby facilitating the generation of the desired fluid flow vortex.

[0055] Figure 5A and Figure 5BOne embodiment of an upper geometric flow aid is shown. In this embodiment, the geometric flow aid is an upper cone 35. As described above, the geometric flow aid includes a geometric shape having a decreasing diameter in a direction toward the center of the first chamber 12. Therefore, the upper cone 35 has a tapered portion 38 having a decreasing diameter toward the center of the first chamber 12. The tapered portion 38 is shown as terminating at a cone tip 38a, but in other embodiments, it is contemplated that the tapered portion may terminate before reaching the tip and will have a flattened end portion. For the geometric flow guide, other shapes besides cones may also be considered. The upper cone 35 also includes a plurality of holes 36 in the perforated section, through which the fluid in the first chamber 12 can enter the upper chamber 22. The holes 36 are configured with a suitable diameter to prevent undissolved powdered matrix agglomerates from entering the upper chamber 22. Finally, the upper cone 35 has a base 37 extending radially from the top edge of the perforated section. The base portion 37 is configured to engage with the support structure 34 of the intermediate cover 31 .

[0056] Figure 6 One embodiment of a lower geometric flow aid is shown. In this embodiment, the geometric flow aid is a bottom cone 25. As described above, the geometric flow aid includes a geometric shape having a decreasing diameter in a direction toward the center of the first chamber 12. Therefore, the bottom cone 25 has a tapered portion 28 having a decreasing diameter toward the center of the first chamber 12. The tapered portion 28 is shown as terminating at a tapered tip 28a, but in other embodiments, it is contemplated that the tapered portion terminates before reaching the tip and will have a flattened end portion. For the geometric flow guide, other shapes besides cones are also contemplated. The bottom cone 25 also has a base edge 29 at the open end of the tapered portion. The base edge 29 is configured to engage with the support structure 24 of the lower cover 21.

[0057] The interior of the first chamber is preferably of generally cylindrical configuration. This further creates a vortex guide for the liquid flow. In addition, the cylinder diameter should complement the input fluid velocity. A first chamber diameter that is too large for a given inflow flow will not support sufficient centrifugal force along its sides to maintain a vortex. An internal diameter that is too small may initially produce excessive turbulence, but no vortex will form, which may result in insufficient mixing. Optimally, it has been found that the diameter of the first chamber is proportional to its height. A height-to-diameter ratio greater than about 2.5:1 will generally not support the generation of sufficiently strong vortices at inflow flow rates of about 1-3 liters / minute. The generally cylindrical shape combined with the inlet velocity and inlet angle is therefore combined to produce the desired vortex.

[0058] Alternatively, other chamber configurations with radial symmetry may also be used for the first chamber housing 22. For example, a spherical shape, a hemispherical shape, a toroidal shape, etc. may be selected.

[0059] Some embodiments may include additional mixing or turbulence components, which may, for example, aid in the mixing process, or provide features that aid in breaking up clumps. Such auxiliary components may include protrusions (e.g., bristles or pegs) extending radially toward the center of the chamber, or protrusions attached to the cone or to the center rod and extending radially outward. In some embodiments, certain mixing auxiliary components may be specifically excluded. For example, mixing components such as propellers, magnets, blades, etc. may be specifically excluded from some embodiments.

[0060] Reference again Figure 3 , the upper inner surface of the first chamber housing 22 can be coupled to the intermediate cover 31 and thus close the first chamber 12 together with the intermediate cover 31. The support structure 34 is preferably integrally molded with the intermediate cover 31 or milled to the intermediate cover 31 to form a boss or lip to support the first chamber 12 and the second chamber 14 and other components for joining the first chamber 12 and the second chamber 14 and allowing fluid flow between the chambers. These components include seals 32, 33, top geometric flow aids (such as upper cones 35), and filter discs 39. The components and parts of the intermediate cover 31 will be referred to below. Figures 8A-8C Describe in more detail.

[0061] The two chambers are preferably adjacent to each other and are separated from each other by an intermediate cover 31 . Figure 1-Figure 3 A preferred embodiment is shown in which the first and second chambers 12, 14 are axially aligned in a watertight seal so that fluid enters the first or lower chamber and moves to the second or upper chamber through an intermediate cover including a circular filter disc 39. In this configuration, seals 32, 33 such as resilient O-rings are used to provide a tight seal between the upper and lower chambers. During manufacturing, chemical A is preferably placed in the first chamber 12 before the filter disc 39 has been put into place.

[0062] The upper chamber housing 42 is also preferably covered by the upper cover 41, thereby enclosing the second chamber 14. The support structure 44 is preferably molded integrally with the upper cover 41 or milled into the upper cover 41 to form a shoulder or lip to be placed on the second chamber housing 42 and to receive other components for engaging the second chamber housing 42 and allowing fluid flow between the second chamber 14 and the outflow port 49. Such components include seals 43, 45, 46, retaining plates 47, and filter discs 48. Figures 9A-9D Describe the components and parts of the upper cover 41 in more detail

[0063] Figure 7The assembly of the lower cover 21 is shown. As shown, the lower cover includes a base portion including the inflow port 16a and a support structure 24 formed therein that receives the bottom cone 25. A seal 23 is positioned around the periphery of the bottom cone 25 and is located on the support structure 24. The seal 23 is preferably an elastic O-ring, but can be a gasket or other sealing device known to those skilled in the art. The seal 23 maintains a watertight connection between the first chamber housing 22 and the lower cover 21.

[0064] In the illustrated embodiment, the lower cover 21 also serves as a base for the support rods 64. Figure 1 In the embodiment of FIG. 1 , two sets of support rods 64 and 65 are shown for separately fixing the first chamber 12 and the second chamber 14. Figure 3 , which shows a single set of rods 64 extending the entire length of the mixing device 10. In other embodiments, it is contemplated that the mixing device 10 is strong enough to stand freely and firmly without any support rods 64, 65.

[0065] Fig. 8A , Figure 8B and Figure 8C The components of the intermediate cover 31 are depicted. Fig. 8A As shown, the intermediate cover 31 includes a base with a central opening. A support structure 34 is located within the central opening, and the support structure 34 is configured to receive the seal 33 thereon. The seal 33 includes a plurality of attachment apertures 33a, each of which is configured to receive a screw 40 therethrough to couple the elements of the intermediate cover 31 together. Figure 8B As shown, the base 37 of the upper cone 35 rests on the support structure 34 and the seal 33. The attachment aperture 37a is aligned with the attachment aperture 33a. Finally, a filter disc 39 is positioned on top of the upper cone 35. The filter disc 39 is fixed to the intermediate cover 31 and fixed to the support structure 34 of the intermediate cover 31 by means of set screws 40 or other attachment means through the attachment apertures 33a and 37a.

[0066] The seal 33 is preferably an elastic O-ring, but can be a gasket or other sealing device known to those skilled in the art. The seal 33 maintains a watertight connection between the upper chamber housing 42 and the intermediate cover 31. The filter disc 39 is preferably made of microporous Porex.TM. plastic (Porex Technologies, Fairburn, GA), but it can also be made of porous polypropylene or polyethylene, glass, wool, micron mesh or any of various other inert materials with suitable compatibility with the solvents and powders used in the device. Preferably, the filter material will have a sufficiently small pore size to prevent the escape of the powdered matrix. In some embodiments, the porosity of the filter is in the range of 30 to 200 microns. For the preferred application described herein, the pore width of the filter is preferably about 90 to 130 microns. The filter disc allows liquid to enter the second chamber, but generally prevents undissolved solids from moving from the first chamber 12 to the second chamber 14. Other undissolved solids captured in the microporous filter are subsequently dissolved by the continuous flow of fluid passing through the filter.

[0067] The first chamber housing 22 and the intermediate cover 31 are provided with a fluid-tight seal 32 by using a resilient O-ring. The first chamber housing 22 may be slidably fitted into an annular recess on the cover 31 or threadedly engaged with a base to couple with the support structure 34. The housing may also be sealed to the cover 31 using adhesives, heat seals, or other means known in the art.

[0068] Fig.9A , Fig. 9B , Fig. 9C and Fig.9D The components of the upper cover 41 are depicted. The view in the accompanying drawings is a bottom perspective view of the upper cover 41. Fig.9A As shown, the seal 46 is placed around the outlet aperture of the outflow port 49 . Fig. 9B Depicted is the placement of a filter disc 48 or outflow filter over the top of the seal 46 and the outlet aperture. Fig. 9C The placement of the second seal 45 on the inner face of the filter disc 48 is depicted. Finally, Fig.9D The placement of a retaining plate 47 covering the seals 45, 46 and filter disc 48 is shown, which is retained on the inner face of the upper cover 41 via engagement holes 41a in the upper cover by attachment elements such as screws 40. The seals 45 and 46 are preferably used to provide a watertight seal between the upper cover 41 and the upper chamber housing 42. The outflow filter 48 is preferably located at least about one-eighth of an inch from the interior surface of the upper cover 41. This provides space for the liquid containing chemicals A and B to pass through the outflow filter 48 and exit through the outflow port 49.

[0069] like Fig.10As shown, the outflow port includes a hose barb connector to which a flexible hose is connected to move fluid from the mixing device 10 to a substrate collector such as a container 50. Fig.11 and Fig.12 As shown, the hose is also preferably fixed to the inflow ports 16a, 16b and can be locked in place by a hose barb connector. In a preferred embodiment, standard flexible laboratory tubing with a diameter large enough so that the tube will cross the neck of the hose barb and small enough so that the tube seals over the hose barb nozzle is used to direct the input fluid flow to the mixing chamber or the output flow to the container 50. The other end of the inflow flexible tube is preferably applied directly to the fluid source. In a preferred culture medium application, the inflow ports 16a, 16b are placed in fluid communication with a distilled deionized water (ddH2O) source having a suitable nozzle, such as the ddH2O tap found in most scientific laboratories. Other tubing materials, nozzle adapters, and pumps may need to be used with other water sources or liquid solvents. As Fig.12 As shown, any of the ports or other passages of the mixing device 10 may include a pressure sensor 17. The pressure sensor may be used to quantify and monitor the pressure in the system and make any changes that may be needed to maintain functionality of the system.

[0070] Flexible pipe that is used to provide communication between the system components can for example be sterilized by autoclave or gamma radiation, and assemble together when manufacturing.Preferably in addition, aseptic receiving container is supplied with equipment.Aseptic receiving container can be glass, plastics or metal, and can be preformed or flexible.In a preferred embodiment, receiving container comprises aseptic flexible bag, for example substrate manager product (Irvine Scientific, Santa Ana, Calif. development).

[0071] In a preferred embodiment, the mixing chamber and cover are made of a non-reactive plastic polymer, such as polycarbonate. Alternatively, the cover and chamber may be molded from other plastics including polysulfone. Other materials include polystyrene, polyethylene, polycarbonate, plexiglass, Lucite, polypropylene, metal alloys or glass. Preferably, the chamber housing 22, 42 will be transparent to enable visual observation of its contents or the progress of the mixing cycle.

[0072] In general, the mixing device 10 is preferably made of a material suitable for a cell culture environment, such as a non-toxic, medical grade plastic or other non-toxic material that will not contaminate the matrix. In current designs, stainless steel can be used, however, there is a need for a single-use system. Therefore, the selected material must be relatively inexpensive while being suitable for a cell culture environment to keep the material and manufacturing costs of a single-use product low.

[0073] In preferred applications, the mixing device is used to prepare tissue culture medium. It is conceivable that the mixing chamber will be provided as a powdered matrix pre-filled with various unit volume sizes. For example, it is conceivable to be suitable for preparing 1 liter (L), 10L, 20L, 50L and up to 100L or larger final tissue culture medium volume mixing chamber size. Increasing the amount of powder in the lower chamber will require increasing the cylinder height and / or diameter to produce a vortex of sufficient size, so as to keep the powder moving in the vortex until it dissolves. In addition, a large size may require a pump located on the inflow line to produce enough inflows to maintain the vortex. It is therefore conceivable that each device is specially designed to supplement the final volume of the product to be prepared.

[0074] Testing has determined that a powder volume greater than about 50% of the chamber volume to which the powdered medium is applied results in poor vortex mixing and inefficient liquid reconstitution. Testing has also determined that during operation of the mixing device disclosed herein, improved reconstitution of the powder in the liquid is achieved by occasionally interrupting the inflow for about five seconds. Interrupting the flow temporarily releases the pressure within the chamber, allowing the powder mass to draw fluid into its interior.

[0075] Additionally, a pressure pulse can be applied to the mixing device to improve reconstitution by temporarily closing the outlet valve at the outflow port. This allows the pressure in the chamber to be increased while the valve is clamped. When the valve is released and the pressure drops rapidly, powder clumps that may have formed in the liquid are pulverized. This pressure pulse can be implemented manually by clamping the outlet tube, or by incorporating a pinch valve into the design at the outflow port.

[0076] The container 50 of precalibration can be used for determining the end point of substrate preparation.Alternately, the liquid pump of predetermined volume can be passed through system, or flow meter / totalizer can be used to monitor the volume of final product.It is envisioned in addition that the final volume of liquid product can be determined by weight.The receiving vessel is placed on the scale and the filling container is up to reaching the final weight of the last product.

[0077] For efficient operation of the device, it is very important that the medium powder is kept relatively dry before use. Hygroscopic powders tend to clump under humid conditions making reconstitution difficult. It is therefore envisaged that commercial products including mixing device systems will have powders packaged under vacuum and / or preferably provided with a desiccant.

[0078] It is also contemplated that the apparatus disclosed herein has numerous other commercial or industrial applications. For example, many liquid medications are prepared at a certain frequency and quantity in hospital pharmacies. Physiological saline solutions, digestive preparations, imaging agents, dyes, sterilizing solutions, and anesthetics are all reconstituted as liquids. Providing pre-weighed aliquots ready for reconstitution such as contemplated by the disclosed technology may provide advantages over the prior art.

[0079] Alternative applications include, but are not limited to, the preparation of pesticides, fertilizers, any of a variety of beverages typically prepared from powders (e.g., milk, iced tea, etc.), which can be reconstituted using the disclosed technology according to some embodiments. It is further contemplated that the liquid solvent employed can be water, alcohol, or other organic matter. The solubility characteristics, solvent to be used, the amount required, and the chemical interaction between the solvent and the reconstitution chemical will be used to provide guidance for the size of the mixing chamber and the selection of component materials.

[0080] Various modifications of this technology can be constructed for different end uses. For example, the figures depict a preferred embodiment in which a first mixing chamber is coaxially aligned below a second chamber and separated by a microporous circular filter disk. In this embodiment, both the upper and lower chambers are cylindrical in shape, and the circular filter disk follows the shape of the chamber housing. As described above, the lower chamber preferably has a generally cylindrical shape and geometric flow aids to promote a sufficiently turbulent rotating fluid velocity.

[0081] However, the upper chamber need not have a cylindrical shape. Other shapes are contemplated for the second chamber and for the microporous filter disc. The second chamber may be rectangular, oval, or substantially spherical. Furthermore, the first and second chambers do not necessarily have to be located on top of each other. It is contemplated that the two chambers may be arranged side by side or remotely from each other and be in fluid communication via silicone, glass, or other conventional tubing.

[0082] Furthermore, it is conceivable that the mixing device comprises only one chamber. Fig.13 100 according to the second embodiment is shown in the figure. In this embodiment, only the first chamber 12 is needed. The powdered substrate and the auxiliary additive such as sodium bicarbonate can be provided to the first chamber 12 together. Therefore, only one chamber is needed to dissolve the solid in the fluid. Therefore, the mixing device 100 includes a lower cover 121 similar to the above-mentioned lower cover 21, a chamber housing 122 and an upper cover 131 similar to the intermediate cover 31 and the upper cover 41. In particular, the upper cover 131 includes an inflow port 116b and an outflow port 149, thereby being used to provide fluid to the chamber 112 and to transport the mixed fluid to the substrate container. Other details related to the mixing device 10 can be similarly applied to the mixing device 100.

[0083] The technology according to some embodiments provides a closed, self-contained mixing system to reconstitute a unit dose of a chemical to a known final liquid volume. The discussion provided above serves to indicate those design features that can be modified to adapt the disclosed device for a wide range of applications. The desire for specific inflow port angles, locations, numbers and diameters, as well as the diameter of the chamber size, fluid pressures, and the need for external turbulence generators are design features that can be easily optimized by one skilled in the art for reconstitution of a given formulation.

[0084] While the preferred embodiments described herein utilize powdered chemicals, it is contemplated that the mixing apparatus is equally applicable to the reconstitution of concentrated liquids or sequential combinations of liquids and powders.

[0085] Thus, some embodiments of the technology disclosed herein provide a method and apparatus for single-step preparation and sterilization (if necessary) of a given chemical. The system is closed, so handling is minimized. All chemicals are pre-weighed, maximizing employee efficiency. The closed system also allows complex sequential or multi-component reconstitution and sterilization processes to be performed in a convenient location without risk of contamination and minimizing changes in the final product due to technician error or batch variation. In addition, the combination of the closed system with a desiccant under vacuum produces a pre-packaged unit that has a relatively long shelf life and improved tolerance to temperature changes compared to the corresponding liquid product.

[0086] The entire contents of US Patent No. 5,470,151 are incorporated herein by reference in their entirety.

[0087] The above description details certain embodiments of the systems, devices, and methods disclosed herein. However, it should be understood that no matter how detailed the foregoing description is, the devices and methods can be implemented in a variety of ways. Also as described above, it should be noted that the use of a specific term in describing certain features or aspects of the present technology should not be considered to mean that the term is redefined herein to be limited to any specific features associated with the term in the features or aspects of the technology. Therefore, the scope of the present disclosure should be interpreted in accordance with the attached claims and any equivalents thereof.

[0088] Those skilled in the art will recognize that various modifications and changes may be made without departing from the scope of the described technology. Such modifications and changes are intended to fall within the scope of the embodiments as defined by the appended claims. Those skilled in the art will also recognize that components included in one embodiment are interchangeable with other embodiments; one or more components from an illustrated embodiment may be included in other illustrated embodiments in any combination. For example, any of the various components described herein and / or depicted in the accompanying drawings may be combined, interchanged, or excluded from other embodiments.

[0089] Regarding the use of any plural and / or singular terms herein, those skilled in the art can appropriately convert the plural to the singular and / or the singular to the plural according to the context and / or application. For the sake of clarity, various singular / plural permutations may be explicitly set forth herein.

[0090] Those skilled in the art will understand that, in general, the terms used herein, and particularly in the appended claims, are generally intended to be "open" terms (e.g., the term "including" should be interpreted as "including but not limited to," the terms "containing" and "having" should be interpreted as "including at least" and "having at least," respectively, the term "comprising" should be interpreted as "including but not limited to," etc.). Those skilled in the art will further understand that if a specific number of an introduced claim statement is intended, such intent will be expressly stated in the claim, and in the absence of such a statement, no such intent is present. For example, as an aid to understanding, the following appended claims may contain the use of the introductory phrases "at least one" and "one or more" to introduce claim statements. However, the use of these phrases should not be interpreted to mean that introducing a claim statement by the indefinite article "a" or "an" will limit any particular claim containing such introduced claim statement to embodiments containing only one such statement, even if the same claim includes the introductory phrases "one or more" or "at least one" and an indefinite article such as "a" or "an." In general, "a" and / or "an" should be interpreted as meaning "at least one" or "one or more"; the same is true for the use of definite articles used to introduce claim recitations. In addition, in cases where a convention similar to "at least one of A, B, and C, etc." is used, generally, such construction is intended to mean that a person skilled in the art will understand the meaning of the convention (e.g., "a system having at least one of A, B, and C" would include but is not limited to systems having only A, only B, only C, A and B, A and C, B and C, and / or A, B, and C, etc.). In those cases where a convention similar to "at least one of A, B, or C, etc." is used, generally, such construction is intended to mean that a person skilled in the art will understand the meaning of the convention (e.g., "a system having at least one of A, B, and C" would include but is not limited to systems having only A, only B, only C, A and B, A and C, B and C, and / or A, B, and C, etc.). Those skilled in the art will further understand that, whether in the specification, claims or drawings, virtually any disjunctive word and / or phrase presenting two or more alternative terms should be understood to contemplate the possibility of including one of the terms, any one of the terms, or both of the terms. For example, the phrase "A or B" will be understood to include the possibility of "A" or "B" or "A and B".

[0091] The technology disclosed herein has a variety of applications, and although specific embodiments of the technology have been described in detail, it will be apparent to those skilled in the art that the disclosed embodiments may be modified in light of the design considerations discussed herein. Therefore, the foregoing description is to be considered illustrative rather than limiting, and the true scope of the invention is that defined by the following claims.

Claims

1. A mixing device for mixing a powder with a liquid, the device comprising: a fluid chamber configured to receive the powder; a first cover at a bottom of the fluid chamber; a first inlet port in the first cover and oriented tangentially relative to an inner wall of the fluid chamber for introducing the liquid into the fluid chamber; a geometric fluid flow aid coupled to the first cover, extending away from the first cover in a direction toward a center of the fluid chamber, and having a constantly decreasing diameter from the first cover to a flattened end portion of the geometric fluid flow aid in a direction toward the center of the fluid chamber; Wherein, the first inflow port and the geometric fluid flow aid are arranged so that the constantly decreasing diameter of the geometric liquid flow aid guides the liquid introduced through the first inflow port into a fluid flow vortex, and the fluid flow vortex is suitable for causing mixing of the powder and the liquid while dispersing the powder and preventing the powder from adhering to or agglomerating at the bottom or top of the fluid chamber.

2. The mixing device according to claim 1, wherein The geometric fluid flow aid is concentric with the inner wall of the fluid chamber.

3. The mixing device according to claim 1, wherein The fluid flow vortex provides a swirling, eddy motion to the fluid and the powder.

4. The mixing device according to claim 1, wherein The geometric fluid flow aid is configured as a single conical flow aid in the fluid chamber.

5. The mixing device of claim 1 further comprising a second cover at the top of the fluid chamber, the second cover comprising a second inflow port and an outflow port.

6. The mixing device according to claim 4, wherein: The first inlet port and the second inlet port are oriented tangentially relative to the fluid flow assist member so as to cause the fluid to establish the fluid flow vortex immediately upon entering the chamber via the first inlet port and the second inlet port.

7. The mixing device of claim 4, further comprising a filter material disposed with the second cover and configured to impede the flow of the powder from the fluid chamber through the outflow port until dissolved in the fluid.

8. The mixing device of claim 4, further comprising a second geometric fluid flow aid coupled to the second cover.

9. The mixing device according to claim 6, wherein: The second geometry fluid flow aid includes a perforated section that allows dissolved powder to flow through the second geometry fluid flow aid while obstructing the flow of the powder through the second geometry fluid flow aid.

10. The mixing device according to claim 9, wherein The second geometric fluid flow aid is located between the tapered section of the second geometric fluid flow aid and the second cover.

11. A method for mixing a powder and a liquid, the method comprising: A mixing device is provided, the mixing device comprising: a fluid chamber configured to receive the powder; a first cover at a bottom of the fluid chamber; a first inlet port in the first cover and oriented tangentially relative to an inner wall of the fluid chamber for introducing the liquid into the fluid chamber; a geometric fluid flow aid coupled to the first cover, extending away from the first cover in a direction toward a center of the fluid chamber, and having a constantly decreasing diameter from the first cover to a flattened end portion of the geometric fluid flow aid in a direction toward the center of the fluid chamber; providing the powder in the fluid chamber; and The liquid is introduced into the fluid chamber via the first inlet port and directed into a fluid flow vortex by the geometrically constantly decreasing diameter of the geometric fluid flow aid, wherein the fluid flow vortex causes mixing of the powder and the liquid while dispersing the powder and preventing the powder from adhering to or agglomerating at the bottom or top of the fluid chamber.

12. The method of claim 11, further comprising removing a substance comprising a powder dissolved in the liquid from the fluid chamber via an outflow port in a second cover positioned at a top of the fluid chamber.

13. The method of claim 12, further comprising introducing additional liquid via a second inlet port positioned in the second cover.

14. The method of claim 11, wherein: The geometric fluid flow aid is a single conical flow aid disposed in the fluid chamber.

15. The method of claim 11, comprising providing the liquid into the fluid chamber via the first inlet port, comprising providing the liquid at a velocity or pressure suitable for interacting with the geometric fluid flow aid, thereby causing the fluid flow vortex to be immediately generated when the fluid is introduced into the fluid chamber.

Images